Red Sea Atlantis II brine pool nitrilase with unique thermostability profile and heavy metal toleranceReport as inadecuate




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BMC Biotechnology

, 16:14

Microbial biotechnology

Abstract

BackgroundNitrilases, which hydrolyze nitriles in a one-step reaction into carboxylic acids and ammonia, gained increasing attention because of the abundance of nitrile compounds in nature and their use in fine chemicals and pharmaceutics. Extreme environments are potential habitats for the isolation and characterization of extremozymes including nitrilases with unique resistant properties. The Red Sea brine pools are characterized by multitude of extreme conditions. The Lower Convective Layer LCL of the Atlantis II Deep Brine Pool in the Red Sea is characterized by elevated temperature 68 °C, high salt concentrations 250 ‰, anoxic conditions and high heavy metal concentrations.

ResultsWe identified and isolated a nitrilase from the Atlantis II Deep Brine Pool in the Red Sea LCL. The isolated 338 amino-acid nitrilase NitraS-ATII is part of a highly conserved operon in different bacterial phyla with indiscernible function. The enzyme was cloned, expressed and purified. Characterization of the purified NitraS-ATII revealed its selectivity towards dinitriles, which suggests a possible industrial application in the synthesis of cyanocarboxylic acids. Moreover, NitraS-ATII showed higher thermal stability compared to a closely related nitrilase, in addition to its observed tolerance towards high concentrations of selected heavy metals.

ConclusionThis enzyme sheds light on evolution of microbes in the Atlantis II Deep LCL to adapt to the diverse extreme environment and can prove to be valuable in bioremediation processes.

KeywordsNitrilase Atlantis II Deep brine pool Red Sea Metagenomics Heavy metals tolerance Thermostability AbbreviationsATIIAtlantis II Deep

BSABovine serum albumin

CDScoding DNA sequence

CTDconductivity, temperature and depth

DTTDithiothreitol

FADFlavin adenine dinucleotide

IPTGIsopropyl β-D-1-thiogalactopyranoside

LBLuria bertani

LCLLower convective layer

MWmolecular weight

OD600Optical density at 600 nm

ORFopen reading frame

PDBProtein data bank

PMSFPhenylmethylesulfonyl fluoride

SDS-PAGESodium dodecyl sulfate-polyacrylamide gel electrophoresis

X-gal5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside

Electronic supplementary materialThe online version of this article doi:10.1186-s12896-016-0244-2 contains supplementary material, which is available to authorized users.

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Author: Sarah A. Sonbol - Ari J. S. Ferreira - Rania Siam

Source: https://link.springer.com/







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